本研究针对农牧业中动物源性细菌携带的可移动耐药基因元件沿着食物链向人类传播日益严重趋势，利用近年在原核细胞基因组中发现的一类新的规律成簇间隔短回文重复CRISPR-Cas系统对外源基因入侵有抵御能力，研究对整合子系统传播耐药性的抑制作用。以本实验室检测的整合子介导的耐药菌株为基础，通过生物信息学分析整合酶、CRISPR-Cas系统基因结构，分子生物学手段构建带特异性识别整合酶位点 CRISPR-Cas 系统及含整合酶和耐药基因盒质粒，等位重组去除CRISPR-Cas 系统各效应基因，分别用检测微生物耐药性、整合效率、Real Time-PCR 检测基因 mRNA 水平变化探索 CRISPR-Cas 系统抑制整合子-耐药基因盒传播的基因机制。本课题从基因水平探索抑制微生物耐药传播问题，为 CRISPR-Cas 系统用于原核生物奠定基础，对控制和解决整合子引起的耐药问题提供实验依据。

Antibiotic-resistant mobile genetic elements along the food chain transmission to humans from animal source bacteria has become a serious trend in Agriculture and animal husbandry.This study is focus on a new clustered regularly,spaced short and palindromic repeat CRISPR-Cas system, which was found having the ability to withstand the invasion of exogenous genes. Based on the detected antibiotic-resistant strains from food source mediated by integron, we analyse the gene structure of CRISPR-Cas system and integrase by bioinformatics, constructe CRISPR-Cas system with specific integrase recognition sites and plasmid containing integrase and resistance gene cassette using molecular biology method,and remove the effects of CRISPR-Cas gene by allelic recombination .Antibiotic-resistance and integration efficiency detection,together with mRNA level detection using real-time PCR are also using in this study.This study aims to explore suppression of microbial atibiotic-resistance spread from gene level and to lay the foundation for the CRISPR-Cas system used in prokaryotes, providing experimental and theoretical basis to controlthe problem of antibiotic-resistance caused by integrons.